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Rabbit anti gfp

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Rabbit anti-GFP is a primary antibody that specifically recognizes green fluorescent protein (GFP). It is designed for use in various immunodetection techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to identify and localize GFP-tagged proteins in biological samples.

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Lab products found in correlation

Mouse anti myc, by Santa Cruz Biotechnology (3 mentions) Rabbit anti h3k4me3, by Abcam (3 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (2 mentions) Rabbit anti flag, by Merck Group (2 mentions) Digoxigenin labeled ribonucleotides, by Roche (2 mentions) Nbt and bcip, by Roche (2 mentions) Peroxidase conjugated anti digoxigenin and tyramide signal amplification tsa, by Thermo Fisher Scientific (2 mentions) Slowfade, by Thermo Fisher Scientific (2 mentions) Spe confocal microscope, by Leica (2 mentions) Dmlb microscope, by Leica (2 mentions) Mouse anti flag, by Merck Group (2 mentions) Mouse anti α tubulin, by Merck Group (2 mentions) Mouse anti ha, by Santa Cruz Biotechnology (2 mentions) Lsm 700 confocal microscope, by Zeiss (2 mentions) Bodipy 493 503, by Thermo Fisher Scientific (1 mentions) A11122, by Thermo Fisher Scientific (1 mentions) Duolink 2 detection kit, by Olink (1 mentions) Abc and dab kits, by Vector Laboratories (1 mentions) Goat anti cpa1, by R&D Systems (1 mentions) Rabbit anti ck19, by Abcam (1 mentions)

30 protocols using rabbit anti gfp

1

Antibody Production and Characterization

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The LD dyes Bodipy493/503 (Invitrogen) and monodansyl pentane (MDH; Abgent) were used at 1 µg/ml and 0.1 mM, respectively. Anti-HA (rat 3F10 monoclonal) antibody was purchased from Roche, anti-PGK1 (mouse) from Life Technologies, anti-GFP (rabbit) from Santa Cruz Biotechnology, Inc., and anti-RFP (mouse) from Abcam. Polyclonal anti-Usa1 antibody was raised against a C-terminal peptide of Usa1 and was previously described (Carvalho et al., 2006 (link)). Recombinant protein fragments were used to raise polyclonal antibodies anti-Dga1 (amino acids 123–272) and anti-Pet10 (amino acids 148–283). All the antibodies were raised in rabbits, and sera anti-Dga1 and anti-Pet10 were affinity purified.
Grippa A., Buxó L., Mora G., Funaya C., Idrissi F.Z., Mancuso F., Gomez R., Muntanyà J., Sabidó E, & Carvalho P. (2015). The seipin complex Fld1/Ldb16 stabilizes ER–lipid droplet contact sites. The Journal of Cell Biology, 211(4), 829-844.
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2

Antibody Validation for Subcellular Imaging

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Anti-Nup210/rabbit (IQ294) purchased from Immunoquest was used for Western blot analysis (1:1,000), while anti-Nup210/rabbit (A301-795A) obtained from Bethyl Laboratories, Inc. was used for immunofluorescence (1:500). mAb414/mouse was purchased from Covance. Anti-MHC antibody/mouse (MF-20) was obtained from Developmental Studies Hybridoma Bank at the University of Iowa. Anti-GFP/rabbit used for Western blotting was obtained from Santa Cruz Biotechnology, Inc. (sc-8384; 1:1,000). Anti-GFP/rabbit used for immunofluorescence was purchased from Invitrogen (A-11122; 1:1,000). Anti-GAPDH/mouse (6C5) was obtained from Abcam (1:2,000). Anti-PDI/mouse used for immunofluorescence was obtained from Abcam (ab5484; 1:1,000) and for Western blot/rabbit from Cell Signaling Technology (mAb #3501; 1:1,000). Anti-BiP/rabbit was purchased from Cell Signaling Technology (mAb #3177). Anti-ATF6/rabbit was obtained from Santa Cruz Biotechnology, Inc. (sc-22799). Caspase-12/rat antibody was obtained from Sigma-Aldrich (#C7611). Anti–caspase-9/rabbit (#9504) and anti–activated caspase 3/rabbit (#9661) was obtained from Cell Signaling Technology. TUDCA dihydrate was obtained from Tokyo Chemical Industry Co.
Gomez-Cavazos J.S, & Hetzer M.W. (2015). The nucleoporin gp210/Nup210 controls muscle differentiation by regulating nuclear envelope/ER homeostasis. The Journal of Cell Biology, 208(6), 671-681.
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3

Gephyrin-GFP Interaction in Neurons

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At 24 h after transfection of hippocampal neurons, PLA was carried out using Duolink II detection kit (Olink Bioscience). Blocking and incubation of primary antibodies and mouse and rabbit secondary probes were performed as described in the immunostaining section. PLA ligation and amplification of PLA dots was performed following the manufacturer's instructions. The following antibodies have been used: mouse anti-gephyrin (clone 3B11) and rabbit anti-GFP (Santa Cruz). Coverslips were mounted on glass slides using Fluoro gel II mounting medium and visualized by fluorescence microscopy.
Dejanovic B., Semtner M., Ebert S., Lamkemeyer T., Neuser F., Lüscher B., Meier J.C, & Schwarz G. (2014). Palmitoylation of Gephyrin Controls Receptor Clustering and Plasticity of GABAergic Synapses. PLoS Biology, 12(7), e1001908.
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4

Quantification of Pancreatic ADM and AFLs

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For quantification of ADM and AFLs, random whole pancreatic sections stained with hematoxylin and eosin H&E were analyzed. For immunohistochemical staining, sections of paraffin embedded pancreata were rehydrated and antigen retrieval was performed using Antigen Unmasking Solution (Vector Laboratories). Overnight incubation with the following primary antibodies was done at 4°C: goat anti-CPA1 (R&D), rabbit anti-CK19 (Abcam), rabbit anti-GFP (Santa Cruz Biotechnology), mouse anti-Ki67 (BD), rabbit anti-P53 (Vector Laboratories), goat anti-Amylase (Santa Cruz Biotechnology), rabbit anti-p44/p42 MAPK (Cell Signaling) and FITC-conjugated DBA-lectin (Vector Laboratories). Biotin-conjugated secondary antibodies were incubated for 1 h at room temperature, following development with ABC and DAB kits (both Vector Laboratories). Nuclear counterstaining was performed using haematoxylin. For immunofluorescence, sections were incubated with fluorophore-conjugated secondary antibody for 1 h at room temperature. Slides were mounted with DAPI hardset antifade mounting medium.
von Figura G., Fahrenkrog-Petersen L., Hidalgo-Sastre A., Hartmann D., Hüser N., Schmid R.M., Hebrok M., Roy N, & Esposito I. (2017). Atypical flat lesions derive from pancreatic acinar cells. Pancreatology : official journal of the International Association of Pancreatology (IAP) ... [et al.], 17(3), 350-353.
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5

Immunoprecipitation of GFP and ATXN3 in HEK293 Cells

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Human embryonic kidney 293 (HEK293) cells were cultured in DMEM, supplemented with 10% FBS, 100U/ml penicillin/streptomycin. Transfections were carried out with lipofectamine 2000 (invitrogen) as described previously (Zeng et al., 2012 (link)). Transfected cells were lysed with 1% Triton X-100 lysis buffer containing 150 mM NaCl, 20 mM Tris/HCl, pH 8.0, 5 mM EDTA, 2 mM N-ethylmaleimide (Life technologies) and complete Mini Protease Inhibitor tablets (Roche). Nuclei and insoluble material were removed by centrifugation. Antigen-antibody complexes were immunoprecipitated with anti-GFP (Invitrogen) or anti-ATXN3 1H9 and protein G beads (Sigma) at 4°C overnight with rotation. After washing three times in lysis buffer, the immunoprecipitates were dissolved in SDS loading buffer and loaded on 10% SDS/PAGE and transferred to polyvinylidene difluoride membranes. The membranes were incubated with the following antibodies: rabbit anti-GFP (1:1000; Santa Cruz biotechnology), rabbit anti-tubulin-α (1:10,000; cell signaling), rabbit anti-flag (1: 5000; Sigma), mouse anti-ATXN3 1H9 (1:1000; Millipore). After incubation with secondary antibodies, blots were developed with blue basic autorad films (GeneMate).
Zeng L., Wang B., Merillat S.A., Minakawa E., Perkins M.D., Ramani B., Tallaksen-Greene S.J., do Carmo Costa M., Albin R.L, & Paulson H.L. (2015). Differential recruitment of UBQLN2 to nuclear inclusions in the polyglutamine diseases HD and SCA3. Neurobiology of disease, 82, 281-288.
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6

Quantifying PRRT2 Protein Expression

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HEK 293 cells were transfected with pEGFP-C1 producing a ∆cPRRT2 fusion protein or wild-type PRRT2 to measure protein expression from the wild-type or mutant genes. Cells were harvested 24 h after transfection. To study the influence of ∆cPRRT2 protein on the wild-type PRRT2 protein, wild-type and mutant plasmids were co-transfected into HEK293 cells with 2 μg of wild-type plasmid and 2 μg, 1.5 μg, 1 μg or 0.5 μg mutant plasmid. EGFP-C1 vector was used to balance the dose to 0, 0.5 μg, 1 μg or 1.5 μg. Cells were homogenized in RIPA buffer with Protease Inhibitor Cocktail (Sigma) and blots were incubated with rabbit anti-GFP (1:1000, Santa Cruz) overnight at 4°C, then goat anti-rabbit IgG-HRP (1:5000, Santa Cruz) at room temperature for 1 hr, then detected with Immobilon Western Chemiluminescent HRP substrate (Millipore).
Ji Z., Su Q., Hu L., Yang Q., Liu C., Xiong J, & Xiong F. (2014). Novel loss-of-function PRRT2 mutation causes paroxysmal kinesigenic dyskinesia in a Han Chinese family. BMC Neurology, 14, 146.
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7

Antibodies and Reagents for Mitochondrial Studies

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The following antibodies were used in this study: rabbit anti-Myo19 (HPA059715, 1:1000 for western blotting) from Sigma-Aldrich; rabbit anti-Mic60 (A2751, 1:3000 for western blotting), anti-Metaxin2 (A7958, 1:1000 for western blotting) and rabbit anti-Sam50 (A3401, 1:3000 for western blotting, 1:400 for PLA assay) from ABclonal; mouse anti-Mic60 (sc-390707, 1:200 for immunofluorescence staining) from Santa Cruz Biotechnology; rabbit anti-GFP (598, 1:1000 for western blotting, 1:200 for immunofluorescence staining) and mouse anti-GFP (MO48-3, 1:200 for immunofluorescence staining) from MBL; anti-mouse (sc-516102, 1:4000) and anti-rabbit (sc-2004, 1:4000) horseradish peroxidase (HRP)-conjugated secondary antibodies from Santa Cruz Biotechnology; Alexa Fluor488- or Alexa Fluor 555-conjugated secondary antibody from ThermoFisher Scientific. For reagents, MitoTrackerTM Red CMXRos (M7512), MitoTrackerTM Green FM (M7514), TMRE (T669), TMRM (T668) and Prolong Diamond Antifade with DAPI (P36962) were purchased from Thermo Fisher Scientific. Rotenone (HY-B1756), FCCP (HY-100410) and 2-Deoxy-D-glucose (HY-13966) were purchased from MedChemExpress.
Shi P., Ren X., Meng J., Kang C., Wu Y., Rong Y., Zhao S., Jiang Z., Liang L., He W., Yin Y., Li X., Liu Y., Huang X., Sun Y., Li B, & Wu C. (2022). Mechanical instability generated by Myosin 19 contributes to mitochondria cristae architecture and OXPHOS. Nature Communications, 13, 2673.
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8

Western Blot Protein Detection

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For western blot analyses, the following antibodies were used: mouse monoclonal anti-GFP (Santa Cruz, 1 : 1000), goat polyclonal anti-dsRED (Santa Cruz, 1 : 1000), rabbit anti-His-tag (Millipore, 1 : 10,000), rabbit anti-mCherry (BioVision, 1 : 5000), and mouse anti-GST tag (Santa Cruz, 1 : 10,000). To immunoprecipitate EGFP-tagged proteins, rabbit anti-GFP (Santa Cruz, 3 μg) or rabbit anti-mCherry (BioVision, 3 μg) was used.
Singh A., Rigatti M., Le A.V., Carlson C.R., Moraru I.I, & Dodge-Kafka K.L. (2015). Analysis of AKAP7γ Dimerization. Journal of Signal Transduction, 2015, 371626.
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9

Western Blot and Immunoprecipitation Assays

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Western blots were carried out on cell lysates separated by 10% SDS-PAGE and performed as previously described [26 (link), 30 (link)]. Immunoprecipitation (IP) assays were carried out using either GFP-trap (ChromoTek), or via protein A Sepharose beads (Life technology), following manufacturer’s protocols with inclusion of protease inhibitor cocktail (Pierce) in cell lysates, as previously described [30 (link)]. For Western blots, anti-epitope tag primary antibodies were used at 1/1000: mouse anti-FLAG M2 (Sigma); rabbit anti-GFP (Santa Cruz); mouse anti-Myc-c (Novus Biologicals); goat anti-human PDGFRA (R&D Systems) [28 , 30 (link)]. gB protein was detected with primary mouse monoclonal antibody to gB [57 ] (a gift from Dr. Britt, UAB) at 1/1000. Rabbit anti-EGFR (Aviva) and mouse anti-beta-actin (Cell Signaling) were used at 1/500. Secondary antibodies for Western blot analysis: anti-mouse, anti-rabbit, or anti-goat IgG/HRP conjugates (Sigma) were used at 1/2000 [28 , 57 , 87 (link)].
El-Hamdi N.S., Choi K.Y, & McGregor A. (2020). Guinea pig cytomegalovirus trimer complex gH/gL/gO uses PDGFRA as universal receptor for cell fusion and entry. Virology, 548, 236-249.
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10

Imaginal Disc Immunostaining Protocol

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Immuno-staining of imaginal discs was according to standard protocols using Hairless anti-A (1:500, from guinea pig) [21 (link)], mouse anti-Cut (1:25) or anti-Wingless (1:25) (developed by G. Rubin and S.M. Cohen, respectively; obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Dept of Biology, Iowa City, IA 52242) and rabbit anti-GFP (1:200, Santa Cruz Biotech, Dallas, USA). Goat secondary antibodies coupled to FITC, Cy3 or Cy5 were from Jackson Immuno-Research (Dianova, Hamburg, Germany). Tissue was mounted in Vectashield (Vector Labs, Eching, Germany), and examined using a BioRad MRC1024 confocal microscope and LaserSharp 2000TM software (Carl Zeiss, Jena, Germany).
Praxenthaler H., Smylla T.K., Nagel A.C., Preiss A, & Maier D. (2015). Generation of New Hairless Alleles by Genomic Engineering at the Hairless Locus in Drosophila melanogaster. PLoS ONE, 10(10), e0140007.
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